Generation of iPS Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

Methods Mol Biol. 2016:1357:57-69. doi: 10.1007/7651_2014_139.

Abstract

Peripheral blood is the easy-to-access, minimally invasive, and the most abundant cell source to use for cell reprogramming. The episomal vector is among the best approaches for generating integration-free induced pluripotent stem (iPS) cells due to its simplicity and affordability. Here we describe the detailed protocol for the efficient generation of integration-free iPS cells from peripheral blood mononuclear cells. With this optimized protocol, one can readily generate hundreds of iPS cell colonies from 1 ml of peripheral blood.

Keywords: Episomal vectors; Hematopoietic cells; Human peripheral blood; Induced pluripotent stem cells; Integration-free; Mononuclear cells; Reprogramming.

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation
  • Cells, Cultured
  • Cellular Reprogramming
  • Gene Expression
  • Genetic Vectors / genetics*
  • Hepatitis B Virus, Woodchuck / genetics
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Leukocytes, Mononuclear / cytology*
  • Plasmids / genetics*
  • Promoter Regions, Genetic / genetics
  • Recombinant Proteins
  • Response Elements / genetics
  • Spleen Focus-Forming Viruses / genetics
  • Transfection
  • bcl-X Protein / genetics
  • bcl-X Protein / physiology

Substances

  • BCL2L1 protein, human
  • Recombinant Proteins
  • bcl-X Protein